title: In vitro fertilization and embryo development in vitro and in vivo in the tiger (Panthera tigris).
abstract: A study was conducted to evaluate the adaptability to the tiger of an in vitro fertilization/embryo culture system previously developed in the domestic cat. In Trial I (July 1989), 10 female tigers were treated with either 2,500 (n = 5) or 5,000 (n = 5) IU eCG i.m. and with 2,000 IU hCG i.m. 84 h later. In Trial II (January 1990), 6 females (5 of which were treated in Trial I) were given 2,500 IU eCG i.m. and 2,000 IU hCG i.m. 84 h later. Twenty-four to twenty-six hours after hCG treatment, all tigers were subjected to laparoscopy, and oocytes were aspirated transabdominally. On the basis of follicular development (follicles greater than or equal to 2 mm in diameter), all females responded to exogenous gonadotropins (range, 6-52 follicles/female). Follicle number and oocyte recovery rate were unaffected (p greater than 0.05) by eCG dose or time of year. A total of 456 oocytes were collected from 468 follicles (97.4% recovery; mean, 28.5 +/- 3.4 oocytes/female). Of these, 378 (82.9%) qualified as mature, 48 (10.5%) as immature, and 30 (6.6%) as degenerate. During Trial I, 8 electroejaculates were collected from 7 male tigers, and in Trial II, 3 semen samples were collected from 3 males. Motile sperm were recovered on each occasion; the overall mean (+/- SEM) ejaculate volume was 7.5 +/- 0.7 ml, the number of motile sperm/ejaculate was 105.9 +/- 20.6 x 10(6), and the percentage of structurally normal sperm/ejaculate was 81.4 +/- 2.0%. After swim-up processing, 0.05 x 10(6) motile sperm were co-cultured with 10 or fewer tiger oocytes in a humidified atmosphere (38 degrees C) of 5% CO2 in air. Of the 358 mature oocytes inseminated, 227 (63.4%) were fertilized. Oocytes from 2 females became contaminated in culture and, therefore, were excluded from embryo cleavage calculations. Of the remaining 195 fertilized oocytes, 187 (95.9%) cleaved to the two-cell stage. No parthenogenetic cleavage was observed in noninseminated control oocytes (n = 20). Eighty-six good-to-excellent-quality two- to four-cell embryos were transferred surgically into the oviducts of 4 of the original oocyte donors in Trial I and 2 females in Trial II. A pregnancy occurred in 1 female in Trial II, and 3 live-born cubs were delivered by Caesarean section 107 days after embryo transfer. Of the 56 cleaved embryos cultured in vitro in Ham's F10 for 72 h, 14 (25.0%) were at the sixteen-cell stage, and 15 (26.8%) were morulae.(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Department of Animal Health, National Zoological Park, Smithsonian Institution, Washington, DC 20008.
authors.forename: A M
authors.initials: AM
authors.lastname: Donoghue
authors.forename: L A
authors.initials: LA
authors.lastname: Johnston
authors.forename: U S
authors.initials: US
authors.lastname: Seal
authors.forename: D L
authors.initials: DL
authors.lastname: Armstrong
authors.forename: R L
authors.initials: RL
authors.lastname: Tilson
authors.forename: P
authors.initials: P
authors.lastname: Wolf
authors.forename: K
authors.initials: K
authors.lastname: Petrini
authors.forename: L G
authors.initials: LG
authors.lastname: Simmons
authors.forename: T
authors.initials: T
authors.lastname: Gross
authors.forename: D E
authors.initials: DE
authors.lastname: Wildt
mesh_terms: D000818:Animals; D002415:Cats; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005314:Embryonic and Fetal Development / Q000187:drug effects / Q000502:physiology*; D005260:Female; D005298:Fertility / Q000187:drug effects; D005307:Fertilization in Vitro / Q000187:drug effects; D008297:Male; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D009866:Oogenesis / Q000187:drug effects; D011247:Pregnancy; D013081:Sperm Motility / Q000187:drug effects / Q000502:physiology; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology
chemical_list: D006063:Chorionic Gonadotropin